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Ku-binding motifs in RAG2, XLF, PAXX and MRI support functional redundancy during V(D)J recombination

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Tadi, Satish | Gesnik, Armelle | Frit, Philippe | Iehl, Florence | Ropars, Virginie | Dingli, Florent | Loew, Damarys | Calsou, Patrick | Callebaut, Isabelle | Charbonnier, Jean-Baptiste | de Villartay, Jean-Pierre

Edité par CCSD ; BioRxiv -

International audience. Abstract The interaction of several partners with Ku through Ku-binding motifs (KBMs) in their sequences governs their enrolment in NHEJ repair complexes. Here, we first established more specifically the function of KBMs in V(D)J recombination as the molecular basis of functional redundancy between XLF and the NHEJ proteins MRI and PAXX. Then, given the functional redundancy between RAG2 and XLF, we explored the hypothesis of a KBM-mediated interaction between RAG2 and Ku. Through sequence alignment and biophysical methods, we identified a KBM at the C-terminus of RAG2 (R2CT) that mediates its interaction with Ku both in vitro and in cellulo . Notably, we showed that R2CT/Ku interaction is independent of the RAG nuclease activity. Finally, we demonstrated that the respective KBMs of RAG2 and XLF support their functional redundancy for V(D)J recombination.

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